BIN overlap confirms transcontinental distribution of pest aphids (Hemiptera: Aphididae)

Muhammad Tayyib Naseem; Muhammad Ashfaq; Arif Muhammad Khan; Akhtar Rasool; Muhammad Asif; Paul D. N. Hebert

doi:10.1371/journal.pone.0220426

Peer Review History

Original SubmissionJuly 11, 2019
23 Oct 2019 Decision Letter - Feng ZHANG, Editor PONE-D-19-19598 BIN overlap confirms transcontinental distribution of pest aphids (Hemiptera: Aphididae) PLOS ONE Dear Muhammad Ashfaq, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. We would appreciate receiving your revised manuscript by Dec. 21th. When you are ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. To enhance the reproducibility of your results, we recommend that if applicable you deposit your laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). This letter should be uploaded as separate file and labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. This file should be uploaded as separate file and labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. This file should be uploaded as separate file and labeled 'Manuscript'. Please note while forming your response, if your article is accepted, you may have the opportunity to make the peer review history publicly available. The record will include editor decision letters (with reviews) and your responses to reviewer comments. If eligible, we will contact you to opt in or out. We look forward to receiving your revised manuscript. Kind regards, Feng ZHANG, Ph.D. Academic Editor PLOS ONE Journal Requirements: When submitting your revision, we need you to address these additional requirements. 1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at http://www.journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and http://www.journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf 2. We note that you have stated that you will provide repository information for your data at acceptance. Should your manuscript be accepted for publication, we will hold it until you provide the relevant accession numbers or DOIs necessary to access your data. If you wish to make changes to your Data Availability statement, please describe these changes in your cover letter and we will update your Data Availability statement to reflect the information you provide. Additional Editor Comments (if provided): [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Partly Reviewer #2: Yes Reviewer #3: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: Yes ****** 3. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: Yes ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: Yes ****** 5. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: Line1-2. Basically the manuscript reports a barcode reference library for some Pakistan aphid species, I don't think it's appropriate to use a title focusing on distribution of pest aphids. Even without a DNA barcoding study, I still can know the geographic distribution patterns of species. Line19-38. The abstract with many numbers may let readers get confused. The authors may keep main results and numbers only. Line43, Line44, Line293. The Aphid Species File is a dynamically updating taxonomic reference, so it's not appropriate to cite a version in 2017. And therefore authors need to check the number of species in related groups. Line165-179. For a DNA barcoding research, the morphologically identification maybe the first step and detail statistics should be placed better in Methods section. Line215. 'across much of Pakistan', based on the sampling map, I think "much of Pakistan" is inappropriate. Line225-230. The discussion here may have logic problem. Specimens could only be identified to a genus and especially subfamily levels can be only due to the authors are not familiar with some aphid groups. If you say the reason is about 'cryptic species complexes', meaning at least the samples can be identified genus level based on sequences. And for the cotton aphid example, the authors cited two relatively old references, do you think the information of 'at least 20 morphologically indistinguishable species' is OK now? One lack of the current version is that, for their own dataset, the authors discuss very little about the 'conflicts between taxonomic assignments and sequences' or 'BIN split' and 'BIN merger' based on biology of related aphids. So the depth of current discussion is not enough. Another problem is, whether the authors did think about the sequence quality in the BOLD system? Sequence with problem may lead to wrong split or merger. The fact is that BOLD sequences have quality problem for many insect groups. At least, the authors need to mention and discuss their opinion about this. Reviewer #2: The manuscript “BIN overlap confirms transcontinental distribution of pest aphids” (PONE-D-19-19598) by Naseem and co-authors analyzes 809 DNA barcodes of aphids from Pakistan using the BOLD workbench. Furthermore, they combined their sequence data with already available DNA barcodes, revealing a broad distribution of pest aphids across six continents. It is obvious that DNA barcoding as well as upcoming metabarcoding approaches using high throughput sequencing technologies will play a more and more important role in order to document and assess biodiversity in the near future. Therefore we need more comprehensive sequence libraries for a correct identification. The topic of this manuscript is interesting and appropriate for PLoS ONE, indicating a relevant application of DNA barcoding in order to identify an economically and ecologically important group of species. However, various concerns remain and have to be corrected before the manuscript is suitable for publication. High-throughput sequencing technologies will play an essential role in modern biodiversity research. However, no aspects of this topic are discussed so far. Furthermore, I miss a discussion of possible pitfalls of using DNA barcodes and mtDNA in general in terms of specimen identification, e.g., the existence of incomplete lineage sorting, heteroplasmy etc. as part of the introduction or discussion. Please find some specific comments made via sticky notes on the PDF file of the manuscript. Reviewer #3: This paper provides valuable data of aphid DNA barcodes. The sampling is limited to Pakistan, but focusses on pest species, increasing the applicative value of the data, especially as most pests have wide distributions. The paper does not contain much scientific novelty besides adding new valuable records to the barcode databases. The paper nicely demonstrates the benefits of BINs in assigning specimens into the same taxonomic units when this is otherwise hampered by misidentifications and taxonomic uncertainties. I recommend the publication after a very minor review. I do not have any major comments, although I find that in some places the authors are close of drawing too strong conclusions based on assumptions. For example, they conclude some issues being because of misidentifications in other works, although they hardly could show this being certainly the case. I recommend carefully checking the sentences at lines 270-274 as the sentences there do not appear fully clear to me. For example: “…are they use widely crop plants…” (perhaps “as they use…”). Similarly. “…their transport on produce fruits…”” (perhaps: “…their transport on produced fruits…”). It is my odd hobby to spot glitches in the references. Here we go: References 24 and 25: One initial perhaps lacking with the author Hebert Reference 48. Kirichenkoa > Kirichenko Lines 458-471: line spacing deviates from that of other parts Reference 89: EXploitation > Expointation (supposedly) Reference 91: Footit > Foottit Disclaimer: I am neither Mr Hebert, nor Mrs Kirichenko, nor Mr Foottit. Additional small glitches: Line 559: The word “Linnean” has a different font size. Figure 3: The name “Sarucallis kahawaluokalani” is not italicized Figure 3: The name “Aphidinae” should not be italicized as being a subfamily-level name. ****** 6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review?** For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: No Reviewer #2: No Reviewer #3: No [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files to be viewed.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email us at figures@plos.org. Please note that Supporting Information files do not need this step. Attachments Attachment Submitted filename: PONE-D-19-19598_R.pdf https://doi.org/10.1371/journal.pone.0220426.r001
Revision 1
4 Nov 2019 Author Response November 4, 2019 PONE-D-19-19598: BIN overlap confirms transcontinental distribution of pest aphids (Hemiptera: Aphididae) Response to Reviewers The comments from the Reviewers are included in italics followed by our response. Reviewer Comments to Author: Reviewer #1: COMMENT: Line1-2. Basically the manuscript reports a barcode reference library for some Pakistan aphid species, I don't think it's appropriate to use a title focusing on distribution of pest aphids. Even without a DNA barcoding study, I still can know the geographic distribution patterns of species. Response: The use of BIN assignments to map the geographic distribution of aphids and other species of pest insects is a key message of our study and this is reflected in the title. We certainly agree with the reviewer that species distributions can be mapped in other ways (e.g. via morphological studies by taxonomists). However, since the taxonomic expertise to correctly identify species is often not available, additional/alternative tools are needed to support species identification. DNA barcoding/BINs meet this need. The utility of the BINs system is further enhanced by the capacity of high-throughput sequencers to support metabarcoding, an approach which allows the species composition of bulk samples to be rapidly assessed. COMMENT: Line19-38. The abstract with many numbers may let readers get confused. The authors may keep main results and numbers only. Response: Several of the less important numbers were removed from the Abstract. COMMENT: Line43, Line44, Line293. The Aphid Species File is a dynamically updating taxonomic reference, so it's not appropriate to cite a version in 2017. And therefore authors need to check the number of species in related groups. Response: Reference [3] has been updated to the format suggested by the database developer. COMMENT: Line165-179. For a DNA barcoding research, the morphologically identification maybe the first step and detail statistics should be placed better in Methods section. Response: Detailed statistics were included in the Results section because DNA barcoding was used to support and facilitate the morphological identifications and finalize the species names. We have mentioned this in the first line of the Results. COMMENT: Line215. 'across much of Pakistan', based on the sampling map, I think "much of Pakistan" is inappropriate. Response: “across much of Pakistan” has been changed to “major agricultural areas of Pakistan” since the areas covered in this study are mostly agricultural lands and plains. COMMENT: Line225-230. The discussion here may have logic problem. Specimens could only be identified to a genus and especially subfamily levels can be only due to the authors are not familiar with some aphid groups. If you say the reason is about 'cryptic species complexes', meaning at least the samples can be identified genus level based on sequences. And for the cotton aphid example, the authors cited two relatively old references, do you think the information of 'at least 20 morphologically indistinguishable species' is OK now? Response: We agree with the reviewer’s comment and have expanded the statement by adding – “or their identification was beyond our expertise”. About cotton aphid (Aphis gossypii): Citations supporting the morphological complexity of A. gossypii are from 2007 (Blackman & Eastop) and 2014 (Favret). These authors are renowned aphid taxonomists and their conclusions on A. gossypii have not been challenged. Based on a more recent report (2014), the statement on the number of species in A. gossypii group has been revised and the old citation [76; Stroyan 1984] has been replaced with a new one (Lagos-Kutz et al. 2014). An old citation on the range of host plants [77, 1955] has been replaced with a more recent report by Singh G, Singh NP, Singh R, Singh G, Singh NP (2014). Food plants of a major agricultural pest Aphis gossypii Glover (Homoptera: Aphididae) from India: An updated checklist. International Journal of Life Sciences Biotechnology and Pharma Research. 2014;3: 1–26. COMMENT: One lack of the current version is that, for their own dataset, the authors discuss very little about the 'conflicts between taxonomic assignments and sequences' or 'BIN split' and 'BIN merger' based on biology of related aphids. So the depth of current discussion is not enough. Response: One species in our dataset was assigned to two BINs (BIN split) and two shared a BIN (BIN merger). These cases of BIN-morphology conflicts (split and merger) have now been discussed (lines 377-380; revised manuscript with track changes). COMMENT: Another problem is, whether the authors did think about the sequence quality in the BOLD system? Sequence with problem may lead to wrong split or merger. The fact is that BOLD sequences have quality problem for many insect groups. At least, the authors need to mention and discuss their opinion about this. Response: BOLD follows stringent quality criteria and sequences not meeting them (<1% ambiguous bases, no stop codon or contamination flag) are not assigned a BIN. This has already been stated in the Data Analysis section of the Methods. Only high quality, validated sequences were included in the current study. BOLD is a public resource and allows users to submit sequences and specimen data without prior permission. However, it does provide quality filters that allow users to exclude invalid and poor quality sequences from any analysis. Reviewer #2: COMMENT: The manuscript “BIN overlap confirms transcontinental distribution of pest aphids” (PONE-D-19-19598) by Naseem and co-authors analyzes 809 DNA barcodes of aphids from Pakistan using the BOLD workbench. Furthermore, they combined their sequence data with already available DNA barcodes, revealing a broad distribution of pest aphids across six continents. It is obvious that DNA barcoding as well as upcoming metabarcoding approaches using high throughput sequencing technologies will play a more and more important role in order to document and assess biodiversity in the near future. Therefore we need more comprehensive sequence libraries for a correct identification. The topic of this manuscript is interesting and appropriate for PLoS ONE, indicating a relevant application of DNA barcoding in order to identify an economically and ecologically important group of species. However, various concerns remain and have to be corrected before the manuscript is suitable for publication. High-throughput sequencing technologies will play an essential role in modern biodiversity research. However, no aspects of this topic are discussed so far. Furthermore, I miss a discussion of possible pitfalls of using DNA barcodes and mtDNA in general in terms of specimen identification, e.g., the existence of incomplete lineage sorting, heteroplasmy etc. as part of the introduction or discussion. Response: We have added a statement on the integration of DNA barcoding with high-throughput sequencing workflows to the Introduction. A discussion point on the potential shortcomings of DNA barcoding (heteroplasmy, incomplete lineage sorting) has been included in the Discussion. Please find some specific comments made via sticky notes on the PDF file of the manuscript. COMMENT: L28: Why 801 and not 809 (see above)? Response: This difference reflects the fact that only sequences meeting quality standards (>500 bp, <1% ambiguous bases, no stop codon or contamination flag) were assigned a BIN. Eight sequences didn’t meet the BIN standard. This has been clarified on lines 128-129 in the Methods section. COMMENT: L41: It would be nice to see some images of the analyzed species. Use the options that open access and PLOS ONE offer! Response: The microscope used for morphological identification of aphids lacked a high quality camera. As a result, most of the images lack the resolution to make morphological characters useful for species assignments. However, as mentioned in the manuscript (line 106), all images taken for the study are accessible on BOLD. COMMENT: L45: Sounds a little bit to drastic; please change. Response: “Attack” was changed to “damage”. COMMENT: L52: Cybertaxonomy per se does not represent an "alternative approach for identification". Instead of this, it focusses on comprehensive descriptions allowing valid identifications (e.g., Organisms, Diversity and Evolution 16: 1-12). Please change this part. Response: Cybertaxonomy and the related reference have been removed. A new reference on the use of protein profiling for species identification has been added. Jayaseelan M, Roesler Uwe R. MALDI-TOF MS Profiling-Advances in species identification of pests, parasites, and vectors. Front Cell Infect Microbiol. 2017;7: 184. COMMENT: L70: Keep in mind that BIN assignments on BOLD are constant¬ly updated as new sequences are added, splitting and/or merging individual BINs in light of new data. Therefore, BINs are not written in stone and can change. Therefore it is important to document the specific date of BIN assignment. Response: The date of the BIN assignment has been documented in the Methods section (line 131). COMMENT: L76: See also PLoS ONE 9(9) e106940. Response: Suggested reference (PLoS ONE 9(9) e106940) has been cited. COMMENT: L113: 94 °C (a gap between the number and °C here and in the following). Response: Corrected. COMMENT: L123: What about the deposition of vouchers? Response: Information on the deposition of vouchers has been added (line 126-128). COMMENT: L134: Think about updating your MEGA-version. The most recent version is 10! Response: We thank the Reviewer for this suggestion. We used MEGA v6 just because it has been well tested. We will consider the newer version for our next study. COMMENT: L223: Please rephrase Response: Sentence has been rephrased to clarify the statement. Reviewer #3: COMMENT: This paper provides valuable data of aphid DNA barcodes. The sampling is limited to Pakistan, but focusses on pest species, increasing the applicative value of the data, especially as most pests have wide distributions. The paper does not contain much scientific novelty besides adding new valuable records to the barcode databases. The paper nicely demonstrates the benefits of BINs in assigning specimens into the same taxonomic units when this is otherwise hampered by misidentifications and taxonomic uncertainties. I recommend the publication after a very minor review. I do not have any major comments, although I find that in some places the authors are close of drawing too strong conclusions based on assumptions. For example, they conclude some issues being because of misidentifications in other works, although they hardly could show this being certainly the case. Response: We thank the reviewer for these valuable comments and suggestions. Although our sampling program was limited to one nation (Pakistan), we were able to extend the analysis to global scale by exploiting the BIN system. Our statements on issues related to misidentification of species are not meant to diminish the value of morphological identification. We simply highlight the difficulties of gaining reliable morphological identifications in areas where the required taxonomic specialists are lacking. COMMENT: I recommend carefully checking the sentences at lines 270-274 as the sentences there do not appear fully clear to me. For example: “…are they use widely crop plants…” (perhaps “as they use…”). Similarly. “…their transport on produce fruits…”” (perhaps: “…their transport on produced fruits…”). Response: These sentences have been corrected. COMMENT: It is my odd hobby to spot glitches in the references. Here we go: References 24 and 25: One initial perhaps lacking with the author Hebert Reference 48. Kirichenkoa > Kirichenko Lines 458-471: line spacing deviates from that of other parts Reference 89: EXploitation > Expointation (supposedly) Reference 91: Footit > Foottit Response: These references and line spacing have been corrected. Exploitation is the original word used by the authors, so it was not changed. COMMENT: Additional small glitches: Line 559: The word “Linnean” has a different font size. Figure 3: The name “Sarucallis kahawaluokalani” is not italicized Figure 3: The name “Aphidinae” should not be italicized as being a subfamily-level name. Response: Corrected. Attachments Attachment Submitted filename: Response-to-Reviewers_PONE-D-19-19598.docx https://doi.org/10.1371/journal.pone.0220426.r002
19 Nov 2019 Decision Letter - Feng ZHANG, Editor PONE-D-19-19598R1 BIN overlap confirms transcontinental distribution of pest aphids (Hemiptera: Aphididae) PLOS ONE Dear Muhammad, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. We would appreciate receiving your revised manuscript by Dec. 19th, 2019. When you are ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. To enhance the reproducibility of your results, we recommend that if applicable you deposit your laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). This letter should be uploaded as separate file and labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. This file should be uploaded as separate file and labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. This file should be uploaded as separate file and labeled 'Manuscript'. Please note while forming your response, if your article is accepted, you may have the opportunity to make the peer review history publicly available. The record will include editor decision letters (with reviews) and your responses to reviewer comments. If eligible, we will contact you to opt in or out. We look forward to receiving your revised manuscript. Kind regards, Feng ZHANG, Ph.D. Academic Editor PLOS ONE [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation. Reviewer #2: (No Response) Reviewer #3: All comments have been addressed ******** 2. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #2: Yes Reviewer #3: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #2: Yes Reviewer #3: Yes ****** 4. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #2: Yes Reviewer #3: Yes ****** 5. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #2: Yes Reviewer #3: Yes ****** 6. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #2: It is nice to see that the authors accepted most comments and/or remarks. Some minor concerns still remain (see sticky notes). After correction, the manuscript should be suitable for publication in PLoS ONE. Reviewer #3: I like that the authors have carefully addressed both the issues that I myself spotted, but also the issues spotted by the other reviewers. I noticed that one of my own comments was with a mistake. I definitely did not mean to change the word "exploitation" to "expointation" in the reference list, but to remove the double capital in the previous word (EXploitation > Exploitation). ****** 7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review?** For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #2: No Reviewer #3: No [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files to be viewed.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email us at figures@plos.org. Please note that Supporting Information files do not need this step. Attachments Attachment Submitted filename: PONE-D-19-19598_R1.pdf https://doi.org/10.1371/journal.pone.0220426.r003
Revision 2
19 Nov 2019 Author Response November 19, 2019 PONE-D-19-19598: BIN overlap confirms transcontinental distribution of pest aphids (Hemiptera: Aphididae). Response to Reviewers The comments from the Reviewers are included in italics followed by our response. Reviewer Comments to Author: Reviewer #2: COMMENT: It is nice to see that the authors accepted most comments and/or remarks. Some minor concerns still remain (see sticky notes). After correction, the manuscript should be suitable for publication in PLoS ONE. Response: All the comments, pointed out through sticky notes on the pdf, have been addressed in the revised version. COMMENT: L137: Which program was used? MUSCLE? You should mention it (inc. giving the reference). Response: Nucleotide sequence alignment was performed using ClustalW. This has been mentioned, with relevant reference, in the revised version. COMMENT: L299: Most DOIs and PMIDs are still missing Response: We have followed the reference style for PLOS ONE. This style does not ask for DOIs and PMIDs when the volume and page numbers are available. Reviewer #3: COMMENT: I like that the authors have carefully addressed both the issues that I myself spotted, but also the issues spotted by the other reviewers. I noticed that one of my own comments was with a mistake. I definitely did not mean to change the word "exploitation" to "expointation" in the reference list, but to remove the double capital in the previous word (EXploitation > Exploitation). Response: The comment has been addressed. Attachments Attachment Submitted filename: Response-to-Reviewers_PONE-D-19-19598R1.docx https://doi.org/10.1371/journal.pone.0220426.r004
25 Nov 2019 Decision Letter - Feng ZHANG, Editor BIN overlap confirms transcontinental distribution of pest aphids (Hemiptera: Aphididae) PONE-D-19-19598R2 Dear Dr. Muhammad Ashfaq, We are pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it complies with all outstanding technical requirements. Within one week, you will receive an e-mail containing information on the amendments required prior to publication. When all required modifications have been addressed, you will receive a formal acceptance letter and your manuscript will proceed to our production department and be scheduled for publication. Shortly after the formal acceptance letter is sent, an invoice for payment will follow. To ensure an efficient production and billing process, please log into Editorial Manager at https://www.editorialmanager.com/pone/, click the "Update My Information" link at the top of the page, and update your user information. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org. If your institution or institutions have a press office, please notify them about your upcoming paper to enable them to help maximize its impact. If they will be preparing press materials for this manuscript, you must inform our press team as soon as possible and no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. With kind regards, Feng ZHANG, Ph.D. Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0220426.r005
Formally Accepted
2 Dec 2019 Acceptance Letter - Feng ZHANG, Editor PONE-D-19-19598R2 BIN overlap confirms transcontinental distribution of pest aphids (Hemiptera: Aphididae) Dear Dr. Ashfaq: I am pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please notify them about your upcoming paper at this point, to enable them to help maximize its impact. If they will be preparing press materials for this manuscript, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org. For any other questions or concerns, please email plosone@plos.org. Thank you for submitting your work to PLOS ONE. With kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Feng ZHANG Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0220426.r006

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